Aquaporin‐1, aquaporin‐3 and aquaporin‐5 differentially modulate cell biophysical and biomechanical properties, impacting cell stiffness and cell–cell adhesion

Aquaporins (AQPs) are transmembrane proteins that facilitate the movement of water, glycerol, and other small solutes across cell membranes. AQP1, AQP3, and AQP5 are overexpressed in cancer, contributing to cancer cell proliferation, migration, angiogenesis, and metastasis. Previously, we showed that silencing AQP3 and/or AQP5 in human pancreatic ductal adenocarcinoma cells induced morphological changes and decreased cell–cell adhesion, with AQP5 modulating cell stiffness and membrane fluidity, suggesting that AQPs can impact tumor progression via modulation of cell biophysical and biomechanical properties. In this study, we overexpressed AQP1, AQP3, or AQP5 individually in human kidney (HEK‐293 T) cells to investigate their individual effects on biological processes, cell morphology, and biomechanical features. After validating AQP expression and function, we evaluated their impact on cell proliferation and migration. Although cell proliferation remained unaffected, AQP5 overexpression enhanced cell migration. Afterwards, we assessed the effect of these AQPs on cell morphological properties through atomic force microscopy (AFM) imaging. AQP1‐, AQP3‐, and AQP5‐overexpressing cells showed no significant changes in cell morphology. AFM‐based force spectroscopy demonstrated that AQP1 overexpression increased both cell stiffness and cell–cell adhesion, AQP3 overexpression enhanced cell stiffness but slightly reduced cell–cell adhesion, whereas AQP5 overexpression increased both cell elasticity and cell–cell adhesion. Thus, AQP1, AQP3, and AQP5 may play a crucial role in promoting tumor growth and invasion by differentially modulating cell stiffness and cell–cell adhesion.